Cytokeratins comprise a diverse group of intermediate filament proteins that are expressed as pairs in both keratinized and non-keratinized epithelial tissue. Cytokeratins play a critical role in differentiation and tissue specialization and function to maintain the overall structural integrity of epithelial cells. Cytokeratin 9 is an unusually large, type I acidic cytokeratin that differentiates human plantar and palmar epidermal cells. Cytokeratin 9 localizes to the suprabasal layers as well as the upper epidermal layers such as the glandular ridges and interridges. The domains of cytokeratin 9 include a head, an α-helical coiled-coil-forming rod, and a tail; cytokeratin 9 shares significant homology with cytokeratin 10. Mutations in the cytokeratin 9 gene correlate with the development of epidermolytic palmoplantar keratoderma (EPPK), an autosomal dominant inherited skin disorder that is characterized by hyperkeratosis of the skin over the palms and soles.
Background References
1. De Rosa, L. et al. 2014. Long-term stability and safety of transgenic cultured epidermal stem cells in gene therapy of junctional epidermolysis bullosa. Stem Cell Reports. 2: 1-8.
2. Rice RH et al. 2013. Distinguishing ichthyoses by protein profiling. PLoS One 8:e75355.
Immunofluorescence analysis of paraffin-embedded rat skin tissue labeling Cytokeratin 9 (HA720133F) and Vimentin (EM0401).
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS. And then probed with the primary antibodies Cytokeratin 9 (HA720133F, green) at 1/200 dilution and Vimentin (EM0401, red) at 1/1,000 dilution overnight at 4 ℃, washed with PBS.
iFluor™ 594 conjugate-Goat anti-Mouse IgG (HA1126) was used as the secondary antibody at 1/1,000 dilution. DAPI was used as nuclear counterstain.
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